The Role of the Lysine-Specific Histone Demethylase 6b, Kdm6b In Synapse Maturation

赖氨酸特异性组蛋白去甲基酶 6b、Kdm6b 在突触成熟中的作用

• 批准号: 10226369
• 负责人: Urann Chan
• 金额: $ 3.85万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10226369
• 关键词: Address; Affect; Animals; Architecture; Binding Proteins; Biochemistry; Biological Models; Brain; Cell Cycle; Cells; Cerebellum; Chromatin; Confocal Microscopy; Cytoplasmic Granules; Data; Deposition; Development; Disease; Electroporation; Elements; Enterobacteria phage P1 Cre recombinase; Enzymes; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Histone H3; Histones; Immunohistochemistry; Impairment; In Vitro; Intellectual functioning disability; Knock-out; Knowledge; LoxP-flanked allele; Lysine; Mediating; Methylation; Modeling; Molecular; Morphology; Mus; Mutate; Mutation; Neurodevelopmental Disorder; Neuronal Differentiation; Neurons; Phenotype; Play; Process; Publishing; RNA immunoprecipitation sequencing; Regulation; Resolution; Rodent; Role; Site; Structure; Synapses; Testing; Time; Transcriptional Regulation; autism spectrum disorder; cell type; chromatin immunoprecipitation; course development; density; experimental study; gene function; gene repression; histone demethylase; in vivo; knock-down; nerve stem cell; neuron development; postsynaptic; prevent; programs; recruit; tool

ABSTRACT The development and maturation of neurons require temporal induction of cell-specific gene expression programs. This is due in part to mechanisms of chromatin regulation that make sure the right genes are expressed and the wrong genes are repressed at any given time. While chromatin mechanisms that regulate neuronal differentiation early in development are well-characterized, the mechanisms in maturing fate-committed neurons are much less understood. One example of chromatin regulation is the trimethylation of lysine 27 on histone H3, which is a mark associated with gene repression and functions early on in development to prevent the expression of genes involved in alternative cell-fate determination. Our lab has shown that H3K27me3 is lost at specific sites during neuronal maturation, suggesting that this mark undergoes dynamic regulation over time. Thus, we raised the possibility that regulation of H3K27me3 by chromatin regulators can underlie neuronal maturation. Taking advantage of a well-defined model of neuronal development, the rodent cerebellum, I will characterize the role of Kdm6b, a lysine-specific demethylase involved in demethylating H3K27me3, in cerebellar maturation. Our lab has shown that loss of Kdm6b in cerebellar granule neurons (CGNs) results in a disruption of a mature gene expression program which includes many synaptic genes. Consistent with this, knockdown of Kdm6b in vivo in CGNs resulted in decreased density of PSD-95, a marker for the postsynapse. Thus, I hypothesize that Kdm6b regulates CGN synapse maturation through the temporal induction of this mature gene expression program. Utilizing a combination of powerful genetic tools and molecular biochemistry, I plan to study how loss of Kdm6b affects CGN synapse maturation in a cell-autonomous fashion in vivo over time, as well as dissect the mechanism by which Kdm6b regulates CGN gene expression. Completion of these two aims will define to what extent can changes in gene expression and chromatin regulation manifest as cellular changes in maturing fate-committed neurons. Additionally, this proposal will elucidate exactly how Kdm6b regulates genes to mediate downstream cellular changes.

摘要 神经元的发育和成熟需要细胞特异性基因表达的时间诱导 程序.这部分是由于染色质调节机制，确保正确的基因是 表达和错误的基因在任何给定的时间被抑制。虽然染色质调节机制 发育早期的神经元分化是很好的特征，成熟的机制是命运决定的。 对神经元的了解要少得多。染色质调节的一个实例是在染色质上赖氨酸27的三甲基化。 组蛋白H3是一种与基因抑制相关的标志物，在发育早期起作用， 参与替代细胞命运决定的基因的表达。我们的实验室已经证明H3 K27 me 3是丢失的 在神经元成熟过程中的特定位点，这表明该标记随着时间的推移进行动态调节。 因此，我们提出了通过染色质调节剂对H3 K27 me 3的调节可能是神经细胞凋亡的基础。 成熟利用神经元发育的明确模型，啮齿动物小脑，我将 描述Kdm 6 b在小脑中的作用，Kdm 6 b是一种赖氨酸特异性脱甲基酶，参与H3 K27 me 3的脱甲基化， 成熟我们的实验室已经表明，小脑颗粒神经元（CGN）中Kdm 6 b的缺失会导致 一个成熟的基因表达程序，其中包括许多突触基因。与此相一致， 在CGN中体内Kdm 6 b导致PSD-95的密度降低，PSD-95是突触后的标志物。所以我 假设Kdm 6 b通过该成熟基因的时间诱导来调节CGN突触成熟 表达式程序利用强大的遗传工具和分子生物化学的结合，我计划研究 随着时间的推移，Kdm 6 b的丢失如何以细胞自主的方式在体内影响CGN突触成熟，以及 探讨Kdm 6 b调控CGN基因表达的机制。实现这两个目标， 定义基因表达和染色质调节的变化在多大程度上可以表现为细胞的变化， 成熟的命运决定神经元。此外，该提案将阐明Kdm 6 b如何调控基因 来调节下游细胞的变化。